PhD defence

Tyrosine-Based Bioconjugations. Strain-Promoted Cycloadditions for Site-Specific Generation of Protein Conjugates

PhD candidate mr. JJ (Jorick) Bruins MSc
Promotor prof.dr. FL (Floris) van Delft
Co-promotor dr.ing. HB (Bauke) Albada
Organisation Wageningen University, Laboratory for Organic Chemistry
Date

Fri 21 February 2020 16:00 to 17:30

Venue Auditorium, building number 362
Generaal Foulkesweg 1
362
6703 BG Wageningen
0317-483592

Summary:

Introduction
Antibodies have been utilized for treatment of cancers and other diseases in either its native form (e.g. AstraZeneca’s Fasenra and Imfinzi), or with potent cytotoxic agents conjugated for targeted therapy (e.g. Pfizer’s Ifixi). We have developed a new method of rapid, site-selective and high-yielding conjugation by oxidizing tyrosine residues expressed at the termini of antibodies.[1]
Goal
By exposing tyrosine residues via a tetra-glycyltyrosine (G4Y) tag, they become prone to selective enzymatic oxidation by mushroom tyrosinase, which converts the phenol moiety to a quinone. The quinone can perform rapid strain-promoted oxidation-controlled cyclooctyne-1,2-quinone cycloaddition (SPOCQ) with bicyclo[6.1.0]nonyne (BCN),[1a, 2] or cyclopropanated trans-cyclooctene (cpTCO),[1b] resulting in a stable conjugation of a selected probe to an antibody with yields of over 95%.


This method was applied to prepare i.e. antibody-drug conjugates (ADCs) and fluorescently labelled antibodies, and is being used to bi-functionalize antibodies with immunologically relevant proteins such as interleukins, cytotoxic agents and others.


References
[1] a) Bruins et. al., Bioconjugate Chem. 2017, 28, 1189-1193; b) Bruins et. al., Chem. Commun., 2018, 54, 7338-7341; c) Bruins et. al., Chem.Eur.J. 2018, 24, 4749 –4756
[2] a) Borrmann et. al., Bioconjugate Chem. 2015, 26, 257-261; b) Dommerholt et. al., Angew. Chem. Int. Ed. 2010, 49, 9422-9425.

Acknowledgements

This project was funded by the Institute for Chemical Immunology (ICI)